Authentication of Rusa timorensis DNA Using Sequencing Assay with Modified Extraction Method

Abstract

A technique to isolate DNA from deer meat was established to authenticate the deer species and was successfully extracted from deer meat. The method was based on the Epicentre MasterPure^TM Complete DNA and RNA Purification Kit extraction kit with slight modification by increasing the volume of Proteinase K and RNase from 5 µL to 20 µL each. The recovery of deer DNA was then analysed by Polymerase Chain Reaction using cytochrome b gene oligonucleotide primers for the deer family, which targeted the mitochondrial DNA. The increasing volume of Proteinase K and RNase resulted in clean DNA and no smearing after performing Polymerase Chain Reaction. Furthermore, the amplified DNA product was extended to sequence assay to authenticate deer species above 95% maximum identification. The result showed that the increasing volume of Proteinase K and RNase offered a high yield of DNA recovery, which provided more than 100 ng per μL and was useful in validating animal-based sources, especially in deer products.